Reagent and method for determining leukocytes and hemoglobin in the blood

ABSTRACT

Leukocytes and hemoglobin in the blood are determined in vitro with a reagent which comprises a ferricyanide ion-free aqueous solution containing a quaternary ammonium ion and cyanide ion in amounts sufficient to stromatolyse erythrocyte and platelet cells and to convert hemoglobin to a chromogen for the determinations.



1. A REAGENT FOR USE IN THE DETERMINATION OF LEUKOCYTES AND HEMOGLOBIN IN THE BLOOD, EHICH COMRPISES A FERRICYANIDE ION-FREE AQUEOUS SOLUTION CONTAINING A QUATERNARY AMMONIUM ION AND CYANIDE ION, SAID QUATERNARY AMMONIUM ION HAVING ATTACHED TO NITROGEN THREE SHORT CHAIN ALKYL GROUP AND ONE LONG CHAIN ALKYL GROUP, AND SAID IONS BEING PRESENT IN AMOUNTS SUFFSUFFICIENT TO STROMATOLYSE ERYTHROCYTE AND PLATELET CELLS AND CONVERT HEMOGOBLIN TO A CHROMOGEN FOR SAID DETERMINATIONS.
 2. A reagent according to claim 1 wherein said quaternary ammonium ion is provided by a quaternary ammonium salt incorporated in said solution in a concentration in the range of about 0.5 to 10 percent by weight of said solution, and said cyanide ion is present in a concentration in the range of about 0.0006 to 0.005 molar.
 3. A reagent according to claim 2 wherein said salt is a trimethyl tetradecyl ammonium halide, sulfate, phosphate, or nitrate, and said cyanide ion is provided by an alkali metal cyanide incorporated in said solution.
 4. A reagent according to claim 1 wherein said short chain alkyl groups each have from 1 to 3 carbon atoms and said long chain alkyl group has from 10 to 14 carbon atoms.
 5. A reagent according to claim 4 wherein said quaternary ammonium ion is provided by a quaternary ammonium salt incorporated in said solution in a concentration in the range of about 0.5 to 10 percent by weight of said solution, and said cyanide ion is provided by an alkali metal cyanide incorporated in said solution in a concentration in the range of about 0.0006 to 0.005 molar.
 6. A reagent according to claim 5 wherein said salt is a halide, sulfate, phosphate, or nitrate.
 7. A reagent according to claim 1 comprising a composition having the following ingredients and relative proportions:
 8. In a method of determining leukocytes and hemoglobin in the blood wherein a reagent is reacted with a blood sample to stromatolyse erythrocyte and platelet cells and to convert hemoglobin to a chromogen for said determinations, the improvement which comprises employing as said reagent a ferricyanide ion-free aqueous solution containing a quaternary ammonium ion and cyanide ion, said quaternary ammonium ion having attached to nitrogen three short chain alkyl groups and one long chain alkyl group. 